2939: Difference between revisions

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confirmed, paul, Administrators
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Latest revision as of 15:27, 26 March 2022

not working--->
PCR from 2931
Gibson mix cloning

2935-F4
TCTAGTCCGCCAAGTCTAGcatatgCTAGCtagagaataaac 

1uM 2935-F1
atcatatgCTAGCtagagaataaactagtattcttctggtccccacagactc
1 uM 2938-F1
                                cttctggtccccacagactcagagagaaccGCCACCATGGACTATAAGGAC
2939-R1
CTATGTAAGCAGCTTGCGGCCGCTATTCCTTTTTCTTTTTTGCCTG
(CAGGCAAAAAAGAAAAAGGAATAGCGGCCGCAAGCTGCTTACATAG)

========
Use Nde1-AflII for cloning into 2895 or 2926
2939-R2
TGTAAGCAGCTTaAGCTATTCCTTTTTCTTTTTTGCCTGG
(CCAGGCAAAAAAGAAAAAGGAATAGCTtAAGCTGCTTACA)

===
Enzyme-free cloning

<s>1uM 2935-F1
atcatatgCTAGCtagagaataaactagtattcttctggtccccacagactc</s>




<s>EFC-F                 CatatgaagctTAATACGACTCACTATAagGagaataaactagtattcttc </s>


PCR insert from  2931
               EFC-F1 CatatgaagctTAATACGACTCACTATAagGagaataaac  
                              EFC-    F2  TCACTATAagGagaataaac tagtattcttctggtcccca

1 uM 2938-F1
                                cttctggtccccacagactcagagagaaccGCCACCATGGACTATAAGGAC




2939-EFC-R     caCTAGTCCCCATCCTCGAGCTATTCCTTTTTCTTTTTTG
              (CAAAAAAGAAAAAGGAATAGCTCGAGGATGGGGACTAGtg)


PCR from vector 2893


EFC-V-F CTCGAGGATGGGGACTAGtg
EFC-V-R (2942-R)
 TGAGTCGTATTAagcttcatatGAGCTCGCGTTGCTGGCG
 (CGCCAGCAACGCGAGCTCatatgaagctTAATACGACTCA) 


digest with DpnI, purify, Gibson mix and transform 10G


RTENOTITLE T7ag-pA-eCas9 

Nde1 HindIII
catatgaagct  
( T7 ag)
TAATACGACTCACTATA agG  agaataaactagtattcttc tggtccccacagactcagagagaacc
GCCACCATGGACTATAAGGACCACGACGGAGACTACAAGGATCATGATATTGATTACAAAGACGATGACGATAAGATGGCCCCAAAGAAGAAGCGGAAGGTCGGTATCCACGGAGTCCCAGCAGCCGACAAGAAGTACAGCATCGGCCTGGACATCGGCACCAACTCTGTGGGCTGGGCCGTGATCACCGACGAGTACAAGGTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCACAGCATCAAGAAGAACCTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAGGCCACCCGGCTGAAGAGAACCGCCAGAAGAAGATACACCAGACGGAAGAACCGGATCTGCTATCTGCAAGAGATCTTCAGCAACGAGATGGCCAAGGTGGACGACAGCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGGAAGAGGATAAGAAGCACGAGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTACCACCTGAGAAAGAAACTGGTGGACAGCACCGACAAGGCCGACCTGCGGCTGATCTATCTGGCCCTGGCCCACATGATCAAGTTCCGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACAGCGACGTGGACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACCCCATCAACGCCAGCGGCGTGGACGCCAAGGCCATCCTGTCTGCCAGACTGAGCAAGAGCAGACGGCTGGAAAATCTGATCGCCCAGCTGCCCGGCGAGAAGAAGAATGGCCTGTTCGGAAACCTGATTGCCCTGAGCCTGGGCCTGACCCCCAACTTCAAGAGCAACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTGAGCAAGGACACCTACGACGACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAGTACGCCGACCTGTTTCTGGCCGCCAAGAACCTGTCCGACGCCATCCTGCTGAGCGACATCCTGAGAGTGAACACCGAGATCACCAAGGCCCCCCTGAGCGCCTCTATGATCAAGAGATACGACGAGCACCACCAGGACCTGACCCTGCTGAAAGCTCTCGTGCGGCAGCAGCTGCCTGAGAAGTACAAAGAGATTTTCTTCGACCAGAGCAAGAACGGCTACGCCGGCTACATTGACGGCGGAGCCAGCCAGGAAGAGTTCTACAAGTTCATCAAGCCCATCCTGGAAAAGATGGACGGCACCGAGGAACTGCTCGTGAAGCTGAACAGAGAGGACCTGCTGCGGAAGCAGCGGACCTTCGACAACGGCAGCATCCCCCACCAGATCCACCTGGGAGAGCTGCACGCCATTCTGCGGCGGCAGGAAGATTTTTACCCATTCCTGAAGGACAACCGGGAAAAGATCGAGAAGATCCTGACCTTCCGCATCCCCTACTACGTGGGCCCTCTGGCCAGGGGAAACAGCAGATTCGCCTGGATGACCAGAAAGAGCGAGGAAACCATCACCCCCTGGAACTTCGAGGAAGTGGTGGACAAGGGCGCTTCCGCCCAGAGCTTCATCGAGCGGATGACCAACTTCGATAAGAACCTGCCCAACGAGAAGGTGCTGCCCAAGCACAGCCTGCTGTACGAGTACTTCACCGTGTATAACGAGCTGACCAAAGTGAAATACGTGACCGAGGGAATGAGAAAGCCCGCCTTCCTGAGCGGCGAGCAGAAAAAGGCCATCGTGGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTGAAGCAGCTGAAAGAGGACTACTTCAAGAAAATCGAGTGCTTCGACTCCGTGGAAATCTCCGGCGTGGAAGATCGGTTCAACGCCTCCCTGGGCACATACCACGATCTGCTGAAAATTATCAAGGACAAGGACTTCCTGGACAATGAGGAAAACGAGGACATTCTGGAAGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAGATGATCGAGGAACGGCTGAAAACCTATGCCCACCTGTTCGACGACAAAGTGATGAAGCAGCTGAAGCGGCGGAGATACACCGGCTGGGGCAGGCTGAGCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGTCCGGCAAGACAATCCTGGATTTCCTGAAGTCCGACGGCTTCGCCAACAGAAACTTCATGCAGCTGATCCACGACGACAGCCTGACCTTTAAAGAGGACATCCAGAAAGCCCAGGTGTCCGGCCAGGGCGATAGCCTGCACGAGCACATTGCCAATCTGGCCGGCAGCCCCGCCATTAAGAAGGGCATCCTGCAGACAGTGAAGGTGGTGGACGAGCTCGTGAAAGTGATGGGCCGGCACAAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAGAACCAGACCACCCAGAAGGGACAGAAGAACAGCCGCGAGAGAATGAAGCGGATCGAAGAGGGCATCAAAGAGCTGGGCAGCCAGATCCTGAAAGAACACCCCGTGGAAAACACCCAGCTGCAGAACGAGAAGCTGTACCTGTACTACCTGCAGAATGGGCGGGATATGTACGTGGACCAGGAACTGGACATCAACCGGCTGTCCGACTACGATGTGGACCATATCGTGCCTCAGAGCTTTCTGGCCGACGACTCCATCGACAACAAGGTGCTGACCAGAAGCGACAAGAACCGGGGCAAGAGCGACAACGTGCCCTCCGAAGAGGTCGTGAAGAAGATGAAGAACTACTGGCGGCAGCTGCTGAACGCCAAGCTGATTACCCAGAGAAAGTTCGACAATCTGACCAAGGCCGAGAGAGGCGGCCTGAGCGAACTGGATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAAACCCGGCAGATCACAAAGCACGTGGCACAGATCCTGGACTCCCGGATGAACACTAAGTACGACGAGAATGACAAGCTGATCCGGGAAGTGAAAGTGATCACCCTGAAGTCCAAGCTGGTGTCCGATTTCCGGAAGGATTTCCAGTTTTACAAAGTGCGCGAGATCAACAACTACCACCACGCCCACGACGCCTACCTGAACGCCGTCGTGGGAACCGCCCTGATCAAAAAGTACCCTGCGCTGGAAAGCGAGTTCGTGTACGGCGACTACAAGGTGTACGACGTGCGGAAGATGATCGCCAAGAGCGAGCAGGAAATCGGCAAGGCTACCGCCAAGTACTTCTTCTACAGCAACATCATGAACTTTTTCAAGACCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGGCGCCTCTGATCGAGACAAACGGCGAAACCGGGGAGATCGTGTGGGATAAGGGCCGGGATTTTGCCACCGTGCGGAAAGTGCTGAGCATGCCCCAAGTGAATATCGTGAAAAAGACCGAGGTGCAGACAGGCGGCTTCAGCAAAGAGTCTATCCTGCCCAAGAGGAACAGCGATAAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAAGAAGTACGGCGGCTTCGACAGCCCCACCGTGGCCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAAAGGGCAAGTCCAAGAAACTGAAGAGTGTGAAAGAGCTGCTGGGGATCACCATCATGGAAAGAAGCAGCTTCGAGAAGAATCCCATCGACTTTCTGGAAGCCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTAAGTACTCCCTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAACTGCAGAAGGGAAACGAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTACCTGGCCAGCCACTATGAGAAGCTGAAGGGCTCCCCCGAGGATAATGAGCAGAAACAGCTGTTTGTGGAACAGCACAAGCACTACCTGGACGAGATCATCGAGCAGATCAGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGGACAAAGTGCTGTCCGCCTACAACAAGCACCGGGATAAGCCCATCAGAGAGCAGGCCGAGAATATCATCCACCTGTTTACCCTGACCAATCTGGGAGCCCCTGCCGCCTTCAAGTACTTTGACACCACCATCGACCGGAAGAGGTACACCAGCACCAAAGAGGTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACACGGATCGACCTGTCTCAGCTGGGAGGCGACAAAAGGCCGGCGGCCACGAAAAAGGCCGGCCAGGCAAAAAAGAAAAAGGAA


TAG

Xho1
CTCGAGGATGGGGAC
TAGtgA
 
GCCGGCC
 Not1
GCGGCCGC
 
GCTCGCTTTCTTGCTGTCCAATTTCTATTAAAGGTTCCTTTGTTCCCTAAGTCCAACTACTAAACTGGGGGATATTATGAAGGGCCTTGAGCATCTGGATTCTGCCTAATAAAAAACATTTATTTTCATT
AAGCTGCTTACATAGAACTCGCGGCGATTG
 



Sph1
GCATGC


CGCCTTAAAATTTTTATTTTATTTTTtCTTTTCTTTTCCGAATCGGATTTTGTTTTTAATATTTCAAAAAAAAAAAAAAAAAAAAAAAAACGCGAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAttaatAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAttaatAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAttaattaa

Xba1
tctaga
acgcgTCGAGGGGAATTAATTCTTGAAGACGAAAGGGCCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGAGCTC 




>210316-33_A11_1FC0DAA038.ab1	1225 3/17/21
2939 seq using VEE-seq-R
NNNNNNNNNNNNNNGNNGCTTNTGNNTAAATGTTTTTTATTAGGCAGAAT
CCAGATGCTCAAGGCCCTTCATAATATCCCCCAGTTTAGTAGTTGGACTT
AGGGAACAAAGGAACCTTTAATAGAAATTGGACAGCAAGAAAGCGAGCGC
GGCCGCGGCCGGCTCACTAGTCCCCATCCTCGAGCTATTCCTTTTTCTTT
TTTGCCTGGCCGGCCTTTTTCGTGGCCGCCGGCCTTTTGTCGCCTCCCAG
CTGAGACAGGTCGATCCGTGTCTCGTACAGGCCGGTGATGCTCTGGTGGA
TCAGGGTGGCGTCCAGCACCTCTTTGGTGCTGGTGTACCTCTTCCGGTCG
ATGGTGGTGTCAAAGTACTTGAAGGCGGCAGGGGCTCCCAGATTGGTCAG
GGTAAACAGGTGGATGATATTCTCGGCCTGCTCTCTGATGGGCTTATCCC
GGTGCTTGTTGTAGGCGGACAGCACTTTGTCCAGATTAGCGTCGGCCAGG
ATCACTCTCTTGGAGAACTCGCTGATCTGCTCGATGATCTCGTCCAGGTA
GTGCTTGTGCTGTTCCACAAACAGCTGTTTCTGCTCATTATCCTCGGGGG
AGCCCTTCAGCTTCTCATAGTGGCTGGCCAGGTACAGGAAGTTCACATAT
TTGGAGGGCAGGGCCAGTTCGTTTCCCTTCTGCAGTTCGCCGGCAGAGGC
CAGCATTCTCTTCCGGCCGTTTTCCAGCTCGAACAGGGAGTACTTAGGCA
GCTTGATGATCAGGTCCTTTTTCACTTCTTTGTAGCCCTTGGCTTCCAGA
AAGTCGATGGGATTCTTCTCGAAGCTGCTTCTTTCCATGATGGTGATCCC
CAGCAGCTCTTTCACACTCTTCAGTTTCTTGGACTTGCCCTTTTCCACTT
TGGCCACCACCAGCACAGAATAGGCCACGGTGGGGCTGTCGAAGCCGCCG
TACTTCTTAGGGTCCCAGTCCTTCTTTCTGGCGATCAGCTTATCGCTGTT
CCTCTTGGGCAGGATAGACTCTTTGCTGAAGCCGCCTGTCTGCACCTCGG
TCTTTTTCACGATATTCACTTGGGGCATGCTCAGCACTTTCCGCACGGTG
GCAAAATCCCGGCCCTTATCCCACACGATCTCCCCGGTTTCGCCGTTTGT
CTCGATCAGAGGCGCCTTCCGGATCTCGCCGTTGGCCAGGGTAATCTCGG
TCTTGAAAAAGTTCATGAAGTTGCC

rev comp
GGCAACTTCATGAACTTTTTCAAGACCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGGCGCCTCTGATCGAGACAAACGGCGAAACCGGGGAGATCGTGTGGGATAAGGGCCGGGATTTTGCCACCGTGCGGAAAGTGCTGAGCATGCCCCAAGTGAATATCGTGAAAAAGACCGAGGTGCAGACAGGCGGCTTCAGCAAAGAGTCTATCCTGCCCAAGAGGAACAGCGATAAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAAGAAGTACGGCGGCTTCGACAGCCCCACCGTGGCCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAAAGGGCAAGTCCAAGAAACTGAAGAGTGTGAAAGAGCTGCTGGGGATCACCATCATGGAAAGAAGCAGCTTCGAGAAGAATCCCATCGACTTTCTGGAAGCCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTAAGTACTCCCTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAACTGCAGAAGGGAAACGAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTACCTGGCCAGCCACTATGAGAAGCTGAAGGGCTCCCCCGAGGATAATGAGCAGAAACAGCTGTTTGTGGAACAGCACAAGCACTACCTGGACGAGATCATCGAGCAGATCAGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGGACAAAGTGCTGTCCGCCTACAACAAGCACCGGGATAAGCCCATCAGAGAGCAGGCCGAGAATATCATCCACCTGTTTACCCTGACCAATCTGGGAGCCCCTGCCGCCTTCAAGTACTTTGACACCACCATCGACCGGAAGAGGTACACCAGCACCAAAGAGGTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACACGGATCGACCTGTCTCAGCTGGGAGGCGACAAAAGGCCGGCGGCCACGAAAAAGGCCGGCCAGGCAAAAAAGAAAAAGGAATAGCTCGAGGATGGGGACTAGTGAGCCGGCCGCGGCCGCGCTCGCTTTCTTGCTGTCCAATTTCTATTAAAGGTTCCTTTGTTCCCTAAGTCCAACTACTAAACTGGGGGATATTATGAAGGGCCTTGAGCATCTGGATTCTGCCTAATAAAAAACATTTANNCANAAGCNNCNNNNNNNNNNNNNN


Score	Expect	Identities	Gaps	Strand
2207 bits(1195)	0.0	1197/1198(99%)	0/1198(0%)	Plus/Plus
Query  2     GCAACTTCATGAACTTTTTCAAGACCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGG  61
             ||||| ||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  3302  GCAACATCATGAACTTTTTCAAGACCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGG  3361

Query  62    CGCCTCTGATCGAGACAAACGGCGAAACCGGGGAGATCGTGTGGGATAAGGGCCGGGATT  121
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  3362  CGCCTCTGATCGAGACAAACGGCGAAACCGGGGAGATCGTGTGGGATAAGGGCCGGGATT  3421

Query  122   TTGCCACCGTGCGGAAAGTGCTGAGCATGCCCCAAGTGAATATCGTGAAAAAGACCGAGG  181
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  3422  TTGCCACCGTGCGGAAAGTGCTGAGCATGCCCCAAGTGAATATCGTGAAAAAGACCGAGG  3481

Query  182   TGCAGACAGGCGGCTTCAGCAAAGAGTCTATCCTGCCCAAGAGGAACAGCGATAAGCTGA  241
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  3482  TGCAGACAGGCGGCTTCAGCAAAGAGTCTATCCTGCCCAAGAGGAACAGCGATAAGCTGA  3541

Query  242   TCGCCAGAAAGAAGGACTGGGACCCTAAGAAGTACGGCGGCTTCGACAGCCCCACCGTGG  301
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  3542  TCGCCAGAAAGAAGGACTGGGACCCTAAGAAGTACGGCGGCTTCGACAGCCCCACCGTGG  3601

Query  302   CCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAAAGGGCAAGTCCAAGAAACTGAAGAGTG  361
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  3602  CCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAAAGGGCAAGTCCAAGAAACTGAAGAGTG  3661

Query  362   TGAAAGAGCTGCTGGGGATCACCATCATGGAAAGAAGCAGCTTCGAGAAGAATCCCATCG  421
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  3662  TGAAAGAGCTGCTGGGGATCACCATCATGGAAAGAAGCAGCTTCGAGAAGAATCCCATCG  3721

Query  422   ACTTTCTGGAAGCCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTA  481
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  3722  ACTTTCTGGAAGCCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTA  3781

Query  482   AGTACTCCCTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAAC  541
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  3782  AGTACTCCCTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAAC  3841

Query  542   TGCAGAAGGGAAACGAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTACCTGGCCA  601
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  3842  TGCAGAAGGGAAACGAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTACCTGGCCA  3901

Query  602   GCCACTATGAGAAGCTGAAGGGCTCCCCCGAGGATAATGAGCAGAAACAGCTGTTTGTGG  661
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  3902  GCCACTATGAGAAGCTGAAGGGCTCCCCCGAGGATAATGAGCAGAAACAGCTGTTTGTGG  3961

Query  662   AACAGCACAAGCACTACCTGGACGAGATCATCGAGCAGATCAGCGAGTTCTCCAAGAGAG  721
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  3962  AACAGCACAAGCACTACCTGGACGAGATCATCGAGCAGATCAGCGAGTTCTCCAAGAGAG  4021

Query  722   TGATCCTGGCCGACGCTAATCTGGACAAAGTGCTGTCCGCCTACAACAAGCACCGGGATA  781
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  4022  TGATCCTGGCCGACGCTAATCTGGACAAAGTGCTGTCCGCCTACAACAAGCACCGGGATA  4081

Query  782   AGCCCATCAGAGAGCAGGCCGAGAATATCATCCACCTGTTTACCCTGACCAATCTGGGAG  841
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  4082  AGCCCATCAGAGAGCAGGCCGAGAATATCATCCACCTGTTTACCCTGACCAATCTGGGAG  4141

Query  842   CCCCTGCCGCCTTCAAGTACTTTGACACCACCATCGACCGGAAGAGGTACACCAGCACCA  901
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  4142  CCCCTGCCGCCTTCAAGTACTTTGACACCACCATCGACCGGAAGAGGTACACCAGCACCA  4201

Query  902   AAGAGGTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACACGGA  961
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  4202  AAGAGGTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACACGGA  4261

Query  962   TCGACCTGTCTCAGCTGGGAGGCGACaaaaggccggcggccacgaaaaaggccggccagg  1021
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  4262  TCGACCTGTCTCAGCTGGGAGGCGACAAAAGGCCGGCGGCCACGAAAAAGGCCGGCCAGG  4321

Query  1022  caaaaaagaaaaaGGAATAGCTCGAGGATGGGGACTAGTGAGCCGGCCGCGGCCGCGCTC  1081
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  4322  CAAAAAAGAAAAAGGAATAGCTCGAGGATGGGGACTAGTGAGCCGGCCGCGGCCGCGCTC  4381

Query  1082  GCTTTCTTGCTGTCCAATTTCTATTAAAGGTTCCTTTGTTCCCTAAGTCCAACTACTAAA  1141
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  4382  GCTTTCTTGCTGTCCAATTTCTATTAAAGGTTCCTTTGTTCCCTAAGTCCAACTACTAAA  4441

Query  1142  CTGGGGGATATTATGAAGGGCCTTGAGCATCTGGATTCTGCCTAATAAAAAACATTTA  1199
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  4442  CTGGGGGATATTATGAAGGGCCTTGAGCATCTGGATTCTGCCTAATAAAAAACATTTA  4499

>210316-33_G11_1FC0DAA039.ab1	1303 3/17/21
2939 seq using  EFC-F1 CatatgaagctTAATACGACTCACTATAagGagaataaac 
good

NNNNTNNNNNNNNNNNACTAGTATTCTTCTGGTCCCCACAGACTCAGAGA
GAACCGCCACCATGGACTATAaGGACCACGACGGAGACTACAAGGATCAT
GATATTGATTACAAAGACGATGACGATAAGATGGCCCCAAAGAAGAAGCG
GAAGGTCGGTATCCACGGAGTCCCAGCAGCCGACAAGAAGTACAGCATCG
GCCTGGACATCGGCACCAACTCTGTGGGCTGGGCCGTGATCACCGACGAG
TACAAGGTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCA
CAGCATCAAGAAGAACCTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAA
CAGCCGAGGCCACCCGGCTGAAGAGAACCGCCAGAAGAAGATACACCAGA
CGGAAGAACCGGATCTGCTATCTGCAAGAGATCTTCAGCAACGAGATGGC
CAAGGTGGACGACAGCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGG
AAGAGGATAAGAAGCACGAGCGGCACCCCATCTTCGGCAACATCGTGGAC
GAGGTGGCCTACCACGAGAAGTACCCCACCATCTACCACCTGAGAAAGAA
ACTGGTGGACAGCACCGACAAGGCCGACCTGCGGCTGATCTATCTGGCCC
TGGCCCACATGATCAAGTTCCGGGGCCACTTCCTGATCGAGGGCGACCTG
AACCCCGACAACAGCGACGTGGACAAGCTGTTCATCCAGCTGGTGCAGAC
CTACAACCAGCTGTTCGAGGAAAACCCCATCAACGCCAGCGGCGTGGACG
CCAAGGCCATCCTGTCTGCCAGACTGAGCAAGAGCAGACGGCTGGAAAAT
CTGATCGCCCAGCTGCCCGGCGAGAAGAAGAATGGCCTGTTCGGAAACCT
GATTGCCCTGAGCCTGGGCCTGACCCCCAACTTCAAGAGCAACTTCGACC
TGGCCGAGGATGCCAAACTGCAGCTGAGCAAGGACACCTACGACGACGAC
CTGGACAACCTGCTGGCCCAGATCGGCGACCAGTACGCCGACCTGTTTCT
GGCCGCCAAGAACCTGTCCGACGCCATCCTGCTGAGCGACATCCTGAAAG
TGAACACCGAGATCACCAGGGCCCCCCTGAACGCCTCTATGATCAGNNAT
ACCAACAAGCACCACCAGGACCTGACCCTGCTGAAAGNNCTCGNGCGGAA
ACNNNTGCCTGAGAANNNCAANNGATTTTCTTNCANNNAANNAGAAACGG
CNACCCCGGTTNCTTNNNNNGNGGNCCNNCCNGAAAAATTCTCNNTNNNN
TGC

Score	Expect	Identities	Gaps	Strand
2132 bits(1154)	0.0	1182/1200(99%)	2/1200(0%)	Plus/Plus
Query  17    ACTAGTATTCTTCTGGTCCCCACAGACTCAGAGAGAACCGCCACCATGGACTATAaGGAC  76
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||| ||||
Sbjct  22    ACTAGTATTCTTCTGGTCCCCACAGACTCAGAGAGAACCGCCACCATGGACTATAAGGAC  81

Query  77    CACGACGGAGACTACAAGGATCATGATATTGATTACAAAGACGATGACGATAAGATGGCC  136
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  82    CACGACGGAGACTACAAGGATCATGATATTGATTACAAAGACGATGACGATAAGATGGCC  141

Query  137   CCAAAGAAGAAGCGGAAGGTCGGTATCCACGGAGTCCCAGCAGCCGACAAGAAGTACAGC  196
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  142   CCAAAGAAGAAGCGGAAGGTCGGTATCCACGGAGTCCCAGCAGCCGACAAGAAGTACAGC  201

Query  197   ATCGGCCTGGACATCGGCACCAACTCTGTGGGCTGGGCCGTGATCACCGACGAGTACAAG  256
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  202   ATCGGCCTGGACATCGGCACCAACTCTGTGGGCTGGGCCGTGATCACCGACGAGTACAAG  261

Query  257   GTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCACAGCATCAAGAAGAAC  316
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  262   GTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCACAGCATCAAGAAGAAC  321

Query  317   CTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAGGCCACCCGGCTGAAGAGA  376
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  322   CTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAGGCCACCCGGCTGAAGAGA  381

Query  377   ACCGCCAGAAGAAGATACACCAGACGGAAGAACCGGATCTGCTATCTGCAAGAGATCTTC  436
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  382   ACCGCCAGAAGAAGATACACCAGACGGAAGAACCGGATCTGCTATCTGCAAGAGATCTTC  441

Query  437   AGCAACGAGATGGCCAAGGTGGACGACAGCTTCTTCCACAGACTGGAAGAGTCCTTCCTG  496
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  442   AGCAACGAGATGGCCAAGGTGGACGACAGCTTCTTCCACAGACTGGAAGAGTCCTTCCTG  501

Query  497   GTGGAAGAGGATAAGAAGCACGAGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTG  556
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  502   GTGGAAGAGGATAAGAAGCACGAGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTG  561

Query  557   GCCTACCACGAGAAGTACCCCACCATCTACCACCTGAGAAAGAAACTGGTGGACAGCACC  616
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  562   GCCTACCACGAGAAGTACCCCACCATCTACCACCTGAGAAAGAAACTGGTGGACAGCACC  621

Query  617   GACAAGGCCGACCTGCGGCTGATCTATCTGGCCCTGGCCCACATGATCAAGTTCCGGGGC  676
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  622   GACAAGGCCGACCTGCGGCTGATCTATCTGGCCCTGGCCCACATGATCAAGTTCCGGGGC  681

Query  677   CACTTCCTGATCGAGGGCGACCTGAACCCCGACAACAGCGACGTGGACAAGCTGTTCATC  736
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  682   CACTTCCTGATCGAGGGCGACCTGAACCCCGACAACAGCGACGTGGACAAGCTGTTCATC  741

Query  737   CAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACCCCATCAACGCCAGCGGCGTG  796
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  742   CAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACCCCATCAACGCCAGCGGCGTG  801

Query  797   GACGCCAAGGCCATCCTGTCTGCCAGACTGAGCAAGAGCAGACGGCTGGAAAATCTGATC  856
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  802   GACGCCAAGGCCATCCTGTCTGCCAGACTGAGCAAGAGCAGACGGCTGGAAAATCTGATC  861

Query  857   GCCCAGCTGCCCGGCGAGAAGAAGAATGGCCTGTTCGGAAACCTGATTGCCCTGAGCCTG  916
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  862   GCCCAGCTGCCCGGCGAGAAGAAGAATGGCCTGTTCGGAAACCTGATTGCCCTGAGCCTG  921

Query  917   GGCCTGACCCCCAACTTCAAGAGCAACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTG  976
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  922   GGCCTGACCCCCAACTTCAAGAGCAACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTG  981

Query  977   AGCAAGGACACCTACGACGACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAGTAC  1036
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  982   AGCAAGGACACCTACGACGACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAGTAC  1041

Query  1037  GCCGACCTGTTTCTGGCCGCCAAGAACCTGTCCGACGCCATCCTGCTGAGCGACATCCTG  1096
             ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  1042  GCCGACCTGTTTCTGGCCGCCAAGAACCTGTCCGACGCCATCCTGCTGAGCGACATCCTG  1101

Query  1097  AAAGTGAACACCGAGATCACCAGGGCCCCCCTGAACGCCTCTATGATCA-GNNATACCAA  1155
             | |||||||||||||||||||| ||||||||||| |||||||||||||| |  ||||  |
Sbjct  1102  AGAGTGAACACCGAGATCACCAAGGCCCCCCTGAGCGCCTCTATGATCAAGAGATACG-A  1160

Query  1156  CAAGCACCACCAGGACCTGACCCTGCTGAAAGNNCTCGNGCGGAAACNNNTGCCTGAGAA  1215
             | ||||||||||||||||||||||||||||||  |||| |||| | |   ||||||||||
Sbjct  1161  CGAGCACCACCAGGACCTGACCCTGCTGAAAGCTCTCGTGCGGCAGCAGCTGCCTGAGAA  1220
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